High-Resolution Melting Analysis (HRMA)—More than just sequence variant screening

Authors

  • Rolf H.A.M. Vossen,

    1. Leiden Genome Technology Center (LGTC), Human and Clincal Genetics, Leiden University Medical Center, Leiden, The Netherlands
    Search for more papers by this author
  • Emmelien Aten,

    1. Leiden Genome Technology Center (LGTC), Human and Clincal Genetics, Leiden University Medical Center, Leiden, The Netherlands
    Search for more papers by this author
  • Anja Roos,

    1. Departments of Nephrology and Clinical Chemistry, Leiden University Medical Center, Leiden, The Netherlands
    Search for more papers by this author
  • Johan T. den Dunnen

    Corresponding author
    1. Leiden Genome Technology Center (LGTC), Human and Clincal Genetics, Leiden University Medical Center, Leiden, The Netherlands
    • Human and Clinical Genetics, Leiden University Medical Center, Einthovenweg 20, 2333CZ Leiden, The Netherlands
    Search for more papers by this author

  • For the Focus Section on HRMA Technology

Abstract

Transition of the double-stranded DNA molecule to its two single strands, DNA denaturation or melting, has been used for many years to study DNA structure and composition. Recent technological advances have improved the potential of this technology, especially to detect variants in the DNA sequence. Sensitivity and specificity were increased significantly by the development of so-called saturating DNA dyes and by improvements in the instrumentation to measure the melting behavior (improved temperature precision combined with increased measurements per time unit and drop in temperature). Melt analysis using these new instruments has been designated high-resolution melting curve analysis (HRM or HRMA). Based on its ease of use, simplicity, flexibility, low cost, nondestructive nature, superb sensitivity, and specificity, HRMA is quickly becoming the tool of choice to screen patients for pathogenic variants. Here we will briefly discuss the latest developments in HRMA and review in particular other applications that have thus far received less attention, including presequence screening, single nucleotide polymorphism (SNP) typing, methylation analysis, quantification (copy number variants and mosaicism), an alternative to gel-electrophoresis and clone characterization. Together, these diverse applications make HRMA a multipurpose technology and a standard tool that should be present in any laboratory studying nucleic acids. Hum Mutat 30:1–7, 2009. © 2009 Wiley-Liss, Inc.

Ancillary